Fig. 4. Expression analysis of Wnt signaling components. Tcf4 expression was examined at the 12- (A), 14- (B,C) and 16-somite (D,E) stages. At the 12-somite stage, Tcf4 expression was not detectable in the diencephalon (A). Expression here was first observed at the 14-somite stage in wild type (B), but not in Shh mutants (C). r5 expression of Tcf4 was observed in both wild-type and Shh mutants at this stage (B,C, arrow). At the 16-somite stage, Tcf4 expression became stronger in the wild-type diencephalon (D), but was still not detectable in the diencephalon of Shh mutants (E). Wnt1 (F,G) and Wnt3a (H,I) were also examined at the 14- and 16-somite stages, respectively. Their expression in the dorsal midline of the diencephalon (*) was maintained in Shh mutants (G,I). Arrowhead indicates the diencephalon-midbrain boundary. Scale bars: 100 µm.

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